High glucose regulates cyclin D1/E of human mesenchymal stem cells through TGF-β1 expression via Ca2+/PKC/MAPKs and PI3K/Akt/mTOR signal pathways

Authors

  • Jung Min Ryu,

    1. Department of Veterinary Physiology, Biotherapy Human Resources Center (BK 21), College of Veterinary Medicine, Chonnam National University, Gwangju, Korea
    Search for more papers by this author
  • Min Young Lee,

    1. Department of Veterinary Physiology, Biotherapy Human Resources Center (BK 21), College of Veterinary Medicine, Chonnam National University, Gwangju, Korea
    Search for more papers by this author
  • Seung Pil Yun,

    1. Department of Veterinary Physiology, Biotherapy Human Resources Center (BK 21), College of Veterinary Medicine, Chonnam National University, Gwangju, Korea
    Search for more papers by this author
  • Ho Jae Han

    Corresponding author
    1. Department of Veterinary Physiology, Biotherapy Human Resources Center (BK 21), College of Veterinary Medicine, Chonnam National University, Gwangju, Korea
    • Department of Veterinary Physiology, College of Veterinary Medicine, Chonnam National University, Gwangju 500-757, Korea.
    Search for more papers by this author

Abstract

The elucidation of factors that support human mesenchymal stem cells (hMSCs) growth has remained unresolved partly because of the reliance of many researchers on ill-defined, proprietary medium formulation. Thus, we investigated the effects of high glucose (D-glucose, 25 mM) on hMSCs proliferation. High glucose significantly increased [3H]-thymidine incorporation and cell-cycle regulatory protein expression levels compared with 5 mM D-glucose or 25 mM L-glucose. In addition, high glucose increased transforming growth factor-β1 (TGF-β1) mRNA and protein expression levels. High glucose-induced cell-cycle regulatory protein expression levels and [3H]-thymidine incorporation, which were inhibited by TGF-β1 siRNA transfection and TGF-β1 neutralizing antibody treatment. High glucose-induced phosphorylation of protein kinase C (PKC), p44/42 mitogen-activated protein kinases (MAPKs), p38 MAPK, Akt, and mammalian target of rapamycin (mTOR) in a time-dependent manner. Pretreatment of PKC inhibitors (staurosporine, 10−6 M; bisindolylmaleimide I, 10−6 M), LY 294002 (PI3 kinase inhibitor, 10−6 M), Akt inhibitor (10−5 M), PD 98059 (p44/42 MAPKs inhibitor, 10−5 M), SB 203580 (p38 MAPK inhibitor, 10−6 M), and rapamycin (mTOR inhibitor, 10−8 M) blocked the high glucose-induced cellular proliferation and TGF-β1 protein expression. In conclusion, high glucose stimulated hMSCs proliferation through TGF-β1 expression via Ca2+/PKC/MAPKs as well as PI3K/Akt/mTOR signal pathways. J. Cell. Physiol. 224:59–70, 2010 © 2010 Wiley-Liss, Inc.

Ancillary